The invention discloses an AFM (atomic force microscope) probe structural parameter-based lateral force calibration method. The method includes the following steps that: the normal elastic coefficient kn and normal sensitivity Sn of a probe to be measured are measured under a standard laboratory environment condition; the cantilever beam length l, cantilever beam thickness t and tip height h of the probe are measured; and the data of kn, Sn, l, t, h are substituted into the calculation formula of a transverse force calibration coefficient alpha, so that the lateral force calibration coefficient of the probe can be estimated. According to the method of the invention, auxiliary gratings and any substrates are not needed, and therefore, costs can be effectively reduced; the method is based on the structural parameters of the probe to be measured, the properties of the probe itself are considered in an emphasized manner, and therefore, the influence of external factors on calibration can be avoided; and calculation is simple and convenient, parameters can be obtained quickly and easily. The method can be widely applied to the transverse force calibration of the AFM probes of rectangular microcantilevers.